Rationale:Dysregulated phagocytic clearance within the synovial microenvironment contributes to persistent inflammation and impairs immune homeostasis in rheumatoid arthritis (RA). Because MerTK-positive macrophages are essential for efferocytosis and inflammation resolution, but are functionally impaired in RA, this study aimed to engineer macrophages capable of enhancing tumor necrosis factor-α (TNF-α) clearance and restoring inflammation-resolving macrophage function. Methods:We generated anti-TNFα chimeric antigen receptor macrophages (CAR-M) by replacing the MerTK antigen-binding domain with a TNF-R1 fragment. The phagocytic and degradative capacity of CAR-M toward soluble TNF-α was assessed, together with activation of Rho GTPases Rac1, Cdc42, and RhoA. Macrophage inflammatory mediator production, phenotypic polarization, SOCS1/3, NF-κB, and MAPK signaling were analyzed. The effects of CAR-M cells were further compared with adalimumab and control treatments under synovial fluid stimulation from patients with arthritis and in mice with collagen-induced arthritis (CIA). Results:Anti-TNFα CAR-M showed markedly enhanced phagocytosis and degradation of soluble TNF-α, accompanied by Rac1, Cdc42, and RhoA activation. CAR activation reduced inflammatory mediator production, including soluble TNF-α, interleukin-1 β (IL-1β), and interleukin-6 (IL-6), while increasing interleukin-10 (IL-10) secretion. This functional shift was associated with polarization toward an inflammation-resolving MerTK+CD206+ phenotype, SOCS1/3 upregulation, and NF-κB and MAPK pathway inhibition. Under stimulation with synovial fluid from arthritis patients, anti-TNFα CAR-M decreased TNF-α and IL-6 levels and drove macrophages toward an M2-like anti-inflammatory state. In CIA mice, local in situ CAR-M cell treatment produced stronger therapeutic effects than adalimumab or other controls, significantly alleviating inflammatory activation, clinical symptoms, and joint pathologies. Conclusions:Anti-TNFα CAR-M exert therapeutic effects via a dual mechanism: direct TNF-α clearance and synovial macrophage reprogramming toward a reparative phenotype. This synthetic biology strategy highlights the potential of precise cell engineering to correct phagocytic deficits and resolve chronic inflammation in RA.
Rationale:The blood-brain barrier (BBB) remains a major obstacle to the delivery of therapeutics for central nervous system (CNS) diseases. Although several BBB-penetrating or BBB-bypassing strategies have been investigated, there remains a need for sustained, controllable delivery routes that can support prolonged CNS exposure of BBB-impermeable agents. Recent anatomical studies have shown direct connections between skull bone marrow and brain, suggesting that the skull may serve as an alternative access route to the brain. Here, we assessed the feasibility of sustained intracalvariosseous infusion (ICO) as skull-to-brain delivery approach for BBB-impermeable molecular and nanoscale agents in mice and rabbits. Methods:Four-week ICO was performed using a species-adapted nano-flow pump-cannula configuration that positioned the cannula terminus within the skull diploic space. A positive comparator was established by positioning the terminus beyond the inner skull cortex. Placement was verified by conventional CT and micro-CT. Brain-associated exposure of BBB-impermeable paclitaxel, antisense oligonucleotides and gold nanoparticles was assessed, along with systemic toxicity, bone marrow-derived immune activation, and neuroinflammation. Results:ICO produced measurable brain-associated exposure of all three model agents in both species. Relative to the positive comparator, ICO achieved the highest relative brain-associated exposure for ASO, reaching approximately 24-28% of comparator levels, whereas PTX and AuNP showed lower but detectable exposure. Four-week ICO was not associated with overt hematological, biochemical, histopathological, or neuroinflammatory abnormalities under the present experimental conditions. Conclusions:These findings support the technical feasibility of ICO-based sustained infusion as a skull-to-brain delivery platform for controlled and prolonged CNS exposure of BBB-impermeable molecular and nanoscale agents through an extracerebral skull compartment.
Rationale:Non-small cell lung cancer (NSCLC) develops a high GSH/GPX4 antioxidant phenotype under persistent oxidative pressure, which suppresses membrane lipid peroxidation and ferroptosis, constituting a core mechanism underlying chemotherapy resistance and suboptimal therapeutic efficacy. Breaking this resistance barrier demands not simply attacking the tumor but disarming the antioxidant defense to intensify oxidative damage and awaken durable antitumor immunity. Methods:A self-assembled nano-prodrug, E-R@ISSL, was engineered by co-assembling a carbamate-linked RGD-modified etoposide (ETP) prodrug and a disulfide-bonded indole derivative conjugated to linoleic acid (LA) at an optimal 1:2 molar ratio. The system thus possesses dual αvβ5/mitochondria targeting with CES2/GSH cascade-responsive drug release. Antitumor activity and mechanisms were evaluated in vitro and in both subcutaneous and orthotopic NSCLC mouse models. Results:E-R@ISSL exhibited efficient αvβ5-mediated internalization and mitochondria-targeted delivery. Upon sequential CES2/GSH-triggered disassembly, E-R@ISSL co-released ETP and LA, reduced the available intracellular GSH pool, and functionally dampened the GSH-dependent GPX4 antioxidant defense. ETP induced DNA double-strand breaks and elevated reactive oxygen species (ROS), while LA expanded the oxidizable lipid pool, synergistically driving lipid peroxidation and ferroptosis. Concurrently, the system activated immunogenic cell death (ICD), promoting dendritic cell maturation and enhancing CD8+ T cell infiltration. In orthotopic NSCLC models, E-R@ISSL significantly suppressed tumor progression, prolonged survival, and demonstrated a favorable safety profile with reduced systemic toxicity compared with free ETP. Conclusions:Together, these findings demonstrate that E-R@ISSL achieved coordinated DNA damage, ferroptosis, and ICD induction through its dual-targeted and sequentially activated co-delivery of ETP and LA, concurrently dismantling the GSH/GPX4 antioxidant defense and intensifying oxidative injury. This nano-prodrug thus represents a viable strategy for overcoming treatment resistance and extending durable antitumor responses in NSCLC.
Background:Three-dimensional (3D) visualization of the renal vasculature plays a crucial role in the pathological assessment and mechanistic investigation of kidney diseases. However, existing 3D imaging approaches remain unable to rapidly and accurately achieve continuous visualization spanning from millimeter-scale arteries to micrometer-scale glomerular capillaries. Here, we developed an innovative strategy, termed EBDE clearing, which enables continuous 3D visualization from the renal artery to the glomerulus, systematically characterizes the spatiotemporal architecture of the entire renal arterial tree and glomeruli in unilateral ureteral obstruction (UUO) and diabetic nephropathy (DN) mouse models. Methods:We combined the vascular tracers of Evans blue and dextran with Ethanol-ECi tissue clearing (EBDE clearing) to achieve the improved labeling of all renal arteries and glomeruli. By further integrating light-sheet microscopy-based 3D imaging with reconstruction and segmentation, we performed high-resolution 3D visualization and quantitative analysis of both renal vascular and glomeruli in normal and diseased kidneys. Furthermore, we investigated, classified, and statistically analyzed the spatiotemporal architecture of the arteries and glomeruli in the kidneys of mice with UUO and DN. Results:Firstly, through quantitative analysis of renal artery diameter, vascular branching, and vascular segment straightness, we found that the arteries of both UUO and DN mice exhibited varying degrees of damage. The renal vascular damage in UUO mice was more severe, manifested by a significant reduction in vessel diameter and the number of vascular branches, as well as an increased in the segment straightness of some arteries; in contrast, the pathological changes in the renal arteries of DN mice were mainly concentrated in vessel diameter and the number of vascular branches. We further classified the glomeruli into three distinct types (M1, M2, and M3) based on their origins, and conducted a quantitative analysis of their 3D spatial distribution within the kidneys of UUO and DN mice, and discovered that in UUO mice, there was a severe reduction in the number and the average volume of the both total and the individual M1-M3 glomeruli. However, in the DN mice, apart from no changes in the total and M1 glomerular number, the other relevant pathological parameters of glomeruli also showed a decrease, although the extent of this decrease was much milder compared to that in the UUO mice. Conclusion:EBDE clearing enables continuous, high-resolution 3D visualization and quantitative analysis from the renal artery to the glomerulus in both normal and diseased intact kidneys. The study not only provides important insights into the renal pathophysiology of UUO and DN, but also establishes a rapid and comprehensive strategy for visualizing structural alterations during disease progression in the kidney and potentially other organs.
Radiopharmaceutical therapy (RPT) plays an important role in modern precision oncology. However, treatment activity is still prescribed by fixed-dose protocols rather than adjusted to planned patient-specific absorbed dose (AD). Individualized dosimetry holds promise for improving treatment planning but is not yet standard. One main reason is that present dosimetry workflows depend on repeated imaging at multiple time points and require kinetic modeling and computational modeling. This review covers recent approaches to make dosimetry more practical and easier to use in everyday RPT workflows. We introduce a theoretical framework in which simplification methods, which help ease the clinical burden of dosimetry, are classified into three related categories. The first two categories simplify the process through direct reduction in the burdens of either the acquisition or processing of data: (1) Methodological simplifications through reduction in the imaging burden, such as reduced- or single-time point (STP) imaging; and (2) Computational automation via artificial intelligence (AI) to automate dosimetry, including image processing, segmentation, registration, and AD estimation. Advanced modeling strategies in the third category improve efficiency through inference, thereby enabling estimation of AD from sparsely sampled data. Reduced imaging played a major role in defining these areas. While all of these have their promise, every category of methods has certain drawbacks. STP techniques are less reliable for heterogeneous tumors, black box models have poor transferability and explainability, and digital twin/physiologically-based pharmacokinetic (DT/PBPK) models lack clinical validation. Fundamental dosimetry, with appropriate validity conditions, could allow for reduced reliance on imaging skills and expertise while still retaining sufficient precision for its intended purposes. Standardization of imaging and multicenter analysis is essential for wider adoption of RPTs. A detailed description of models and their effectiveness would be helpful in the clinical application of dosimetry.
Glioblastoma (GBM) has a high rate of post-surgical recurrence, which can be attributed to residual tumor cells as well as to the disruption of the tumor microenvironment (TME) caused by surgery. Here, we define the postoperative cavity microenvironment (POCM) as a pathological niche with distinct spatial and temporal boundaries, and present a list of markers that outline this environment, including low pH, high levels of reactive oxygen species (ROS), high levels of glutathione (GSH), overexpression of matrix metalloproteinases, and tumor tissue mechanical instability. Engineered nanomedicine is evolving from simple drug-carrying entities to the design of smart responsive materials that can be actively tuned to modulate the POCM. In this review, we outline the working mechanisms and main features of different classes of smart responsive materials, which not only release therapeutic agents but also provide spatiotemporal control over drug delivery, enhance penetration into residual tumor tissue, and trigger immune activation at the right time and place. We discuss how local delivery of nanogel-based platforms in combination with systemic immunotherapy and fine-tuning by means of 3D bioprinting and/or microfluidic chip-based platforms can provide effective tools for reducing post-operative GBM recurrence rates.
Colon-targeted delivery of 5-aminosalicylic acid (5-ASA) offers an effective strategy for treating inflammatory bowel disease (IBD) while minimizing systemic side effects. However, conventional oral 5-ASA formulations suffer from premature drug release and poor colon specificity. To overcome these limitations, we developed a biodegradable, prebiotic, and immunomodulatory inulin nanoparticle (INP) platform enabling microbial enzyme-responsive and site-specific release of 5-ASA in the colon. Methods:INP was fabricated via a mild nanoprecipitation method and loaded with 5-ASA (5-ASA@INP). Physicochemical characteristics, drug-loading performance, and enzyme-responsive sequential release profiles under simulated gastrointestinal fluids were evaluated. Antioxidant and anti-inflammatory effects were assessed in vitro. Biodistribution of DiR-labeled INPs was analyzed to demonstrate colon-targeting efficacy. Therapeutic efficacy, inflammatory cytokine expression, Treg responses in the colon and gut-associated lymphoid tissues, and microbiota-related effects, including short-chain fatty acid (SCFA) production, were evaluated in a colitis mouse model. Results:5-ASA@INP showed 97.02% drug-loading efficiency. The cumulative release of 5-ASA was 7.6% in simulated gastric fluid, 18.7% in simulated intestinal fluid, and 99.2% in simulated colonic fluid. In contrast, only 35.6% of 5-ASA was released under enzyme-free simulated colonic conditions. Compared with free 5-ASA, 5-ASA@INP showed enhanced antioxidant and anti-inflammatory effects in vitro. In vivo, INPs preferentially accumulated in the colon with minimal off-target distribution. In colitic mice, 5-ASA@INP significantly attenuated body weight loss (p = 0.0067) and colon shortening (p = 0.0002). Histological analysis showed that the colonic tissue damage was minimized and the inflammation scores were reduced (p < 0.001). Treatment increased the number of FOXP3+ cells in the colonic tissue and suppressed the expression of IL-17A, TNF-α, and IL-1β. The gut microbial profile was restored toward that of the normal group and fecal SCFA levels were increased. Conclusions:This study demonstrates that the INP platform enables microbial enzyme-responsive, colon-targeted delivery of 5-ASA while complementarily harnessing the intrinsic prebiotic and immunomodulatory properties of inulin. This dual-function nanoplatform provides a promising strategy for efficient oral treatment of IBD.
Background:Identifying tumor selective targets is critical for the development of precision diagnostic and therapeutic agents in oncology. Despite advances in precision oncology elsewhere, there are no FDA-approved hepatocellular carcinoma (HCC) antigen-selective antibody-drug conjugates or radiopharmaceuticals. This study establishes an integrated workflow to identify HCC-enriched plasma membrane targets and assess their suitability for targeted molecular imaging as foundational candidates for future radiopharmaceutical therapy. Methods:Bulk RNA sequencing (371 tumors), single cell RNA sequencing (34 HCC cases), and a normal liver dataset were analyzed to identify HCC-enriched plasma membrane targets. Candidate molecules were examined on HCC and normal tissue microarrays (TMAs) and further evaluated in liver cancer cell lines by quantitative PCR, Western blot, and flow cytometry. Selected targets were then tested in mouse models of liver cancer using antibody-based positron emission tomography (immunoPET) to assess in vivo target engagement and distribution. Results:Integrated transcriptomic analysis identified several tumor plasma membrane molecules with strong tumor enrichment, including GPC3, MUC13, TSPAN8, MET, and EGFR. TMAs confirmed prominent membrane expression in HCC with little signal in normal organs. Combinations of four prioritized markers captured up to 88.5% of patient tumors. Antibody-based immunoPET agents directed against prioritized targets demonstrated specific tumor accumulation in vivo, and signal intensity correlated with membrane staining by immunohistochemistry. These results highlight the potential of these markers to serve as diagnostic tools and promising candidate platforms for therapeutic development, demonstrating the utility of this approach to identify novel oncology targets. Conclusions:This study establishes a multimodal framework integrating transcriptomic predictions and experimental protein validation to identify HCC targets for molecular imaging and radiopharmaceutical development. This translational blueprint successfully advances precision diagnostic and theranostic-ready agents for this disease for HCC, and a similar approach may be useful for identifying and validating targets in other malignancies.
Background:The αvβ6 and αvβ8 integrins are upregulated in many solid-tumors and drive local activation of transforming growth factor-β (TGFβ), promoting immune evasion and resistance to immune checkpoint blockade. The chromogranin A-derived peptide 4Δ targets the RGD-binding site of αvβ6/αvβ8 and inhibits integrin-dependent TGFβ-activation. We investigated whether 4Δ-derived peptibodies can deliver cytotoxic drugs to cancer cells and enhance immune checkpoint inhibitors (ICIs) activity. Methods:Peptide 4Δ was genetically fused to the Fc domains of murine and human IgG1 to generate the peptibodies 4ΔmFc and 4ΔhFc. Integrin-binding properties and inhibition of TGFβ activation were characterized using biochemical and cell-based assays. Peptibody internalization and lysosomal trafficking were analyzed by live-cell/confocal microscopy. Cytotoxic activity of peptibodies complexed with anti-Fc antibodies coupled to anticancer drugs was evaluated using αvβ6/αvβ8-positive and -negative cancer cells. Pharmacokinetics and antitumor activity of 4ΔmFc, alone or in combination with an anti-PD-L1 antibody, were evaluated in murine fibrosarcoma and mammary carcinoma models. Results:4ΔmFc and 4ΔhFc bound αvβ6 and αvβ8 with sub-nanomolar affinity. Both compounds selectively recognized αvβ6/αvβ8-positive tumor cells, as well as human pancreatic, lung, and colon carcinomas sections. 4ΔmFc and 4ΔhFc efficiently inhibited TGFβ activation, underwent efficient internalization, and trafficked to lysosomes. 4ΔhFc enabled delivery of cytotoxic payloads to αvβ6/αvβ8-positive cells, including MMAE, MMAF, DM1, PBD, or DX8951. 4ΔmFc delayed the growth of fibrosarcomas and improved mice survival in the mammary adenocarcinoma model when combined with an anti-PD-L1 mAb (immune checkpoint inhibitor), without overt toxicity. Conclusion:These peptibodies represent a dual-selective αvβ6/αvβ8-targeting platform that couples potent blockade of integrin-dependent TGFβ activation with efficient, selective delivery of cytotoxic payloads to cancer cells. These properties, together with the observed synergism with anti-PD-L1 mAbs, suggest their potential use as ligands for delivering cytotoxic agents to tumors, while concomitantly modulating the TGFβ-driven immunosuppressive microenvironment, either alone or in combination with ICIs.
Rationale:Fungal keratitis remains clinically challenging due to limited drug bioavailability, frequent dosing, reactive oxygen species (ROS)-mediated stromal damage, perforation risk, and bacterial coinfection. In the study, we developed a feedback-regulated multifunctional corneal micropatch (MFCP) via 3D printing for the treatment of fungal keratitis. Methods:The dual-network hydrogel micropatch comprises an ROS-responsive hydrogel (RRH), voriconazole-loaded F127DA micelles (VCZ-F127DA), and gatifloxacin-loaded cerium metal-organic frameworks (GAT-MOFs). We characterized its physicochemical properties and evaluated its biocompatibility, corneal healing capacity, ROS scavenging activity, and antimicrobial performance. Histological staining, cytokine assays, and transcriptomic sequencing were used to assess its therapeutic efficacy in a mouse model of fungal keratitis. Results:MFCP exhibits tunable curvature, high transparency, and mechanical properties matching those of the cornea, along with good biocompatibility. High ROS level can accelerate breakdown of the micropatch and promote drug release, generating strong synergistic antifungal and antibacterial activities. The released GAT-MOFs scavenge excessive ROS, forming a negative feedback loop. In the mouse model of fungal keratitis, MFCP mitigates corneal infection, lowers proinflammatory cytokines, and accelerates tissue repair, with therapeutic efficacy superior to that of voriconazole eye drops. Transcriptomic analysis reveals suppression of NF-κB-mediated inflammatory signaling and upregulation of corneal regenerative pathways after treatment with MFCP. Conclusion:These findings demonstrate that MFCP represents a viable therapeutic option for fungal keratitis.
Rationale:A combination of gemcitabine (GEM)-based chemotherapy and an immune checkpoint inhibitor is the standard treatment for patients with advanced intrahepatic cholangiocarcinoma (iCCA). However, 70% of patients develop progressive disease following GEM treatment, highlighting the urgent need for therapeutic strategies targeting GEM-resistant (GR) disease. Methods:We established GR iCCA sublines and identified 9 upregulated genes by RNA-sequencing and cDNA microarray. Among the 9 genes, we demonstrated that VTCN1 expression improved GR. The enhanced effect of VTCN1 on GR was validated in a spontaneous rat iCCA model, a xenograft mouse model, an orthotopic mouse model, and iCCA specimens. A phospho-kinase array was used to identify the downstream effector of membrane and soluble VTCN1 (sVTCN1). Results:VTCN1 expression was significantly elevated in GEM non-responders and independently predicted poor progression-free survival (HR = 2.038, P = 0.035). Plasma sVTCN1 levels correlated with tumor VTCN1 expression (r = 0.8752, P = 0.002) and tumor burden (r = 0.7696, P < 0.0001). Mechanistically, membrane-bound VTCN1 directly interacted with integrin β1 to enhance FAK phosphorylation, while sVTCN1 bound EGFR to create a positive feedback loop increasing EGFR phosphorylation. Both pathways converged on Src family kinase activation (Y419 phosphorylation) to drive GR. The VTCN1-Src axis was associated with upregulation of cytosolic 5'-nucleotidase 3 (NT5C3) expression to promote GEM resistance. VTCN1 depletion, Src depletion, or Src inhibition with dasatinib restored GEM sensitivity. Combining VTCN1 antibody or dasatinib with GEM significantly suppressed resistant tumor growth and enhanced CD8+ T cell infiltration in vivo. Conclusions:Membrane and soluble VTCN1 drive GR through dual pathways converging on Src activation. VTCN1 represents both a prognostic biomarker and therapeutic target, with Src inhibition providing a rational strategy to overcome chemotherapy resistance in iCCA.
Rationale:Lutetium-177 has demonstrated clinical success, particularly in neuroendocrine and prostate cancers, but disease recurrence and progression remain frequent. Terbium-161 offers improved therapeutic potential by delivering higher localized radiation doses. We have previously demonstrated preclinical efficacy of the CD44v6-targeted radiopharmaceutical [177Lu]Lu-AKIR001, which is currently under clinical investigation for multiple malignancies (NCT06639191). The present study explored the terbium-161-labeled analogue preclinically, focusing on pancreatic ductal adenocarcinoma, with early proof-of-concept investigations in squamous cell carcinoma, both being highly aggressive and treatment-resistant malignancies. Methods:Radioligand uptake was evaluated in vitro in squamous cell carcinoma and pancreatic ductal adenocarcinoma cell lines. Ex vivo biodistribution and dosimetry estimations confirmed a selective tumor uptake of [161Tb]Tb-AKIR001 before therapeutic efficacy was evaluated in both squamous cell carcinoma and pancreatic ductal adenocarcinoma xenograft models. Mice were administered [161Tb]Tb-AKIR001 of activities ranging from 4 to 10 MBq and compared to untreated controls. Potential adverse effects were evaluated by blood sampling, monitoring of body weights, and histopathology. Results:Specific, high-affinity binding of [161Tb]Tb-AKIR001 to CD44v6-positive cells was verified in vitro. Tumor uptake in vivo exceeded 30% injected activity per gram of tissue in the A431 model at 96 h post-injection. Peak tumor uptake in the BxPC3 model exceeded 150% injected activity per gram of tissue, with a mean absorbed dose to tumor of 17 Gy/MBq. Activity-dependent antitumor effects were observed in both xenograft models, accompanied by mild to moderate, transient hematologic effects that were significantly mitigated by activity fractionation. Histopathological evaluation revealed no treatment-related tissue damage in kidney, liver or spleen. Conclusion:[161Tb]Tb-AKIR001 demonstrated favorable biodistribution and dosimetry profiles, together with encouraging therapeutic effects without signs of severe toxicity. Compared to [177Lu]Lu-AKIR001, [161Tb]Tb-AKIR001 may offer added advantages in certain therapeutic contexts, and our results highlight [161Tb]Tb-AKIR001 as a promising candidate for further development in targeted radionuclide therapy.
Rationale:Severe acute pancreatitis (SAP) is a life-threatening disease with substantial mortality, primarily resulting from pancreatic microcirculatory failure that initiates multiple organ dysfunction syndrome (MODS). While early microvascular injury is widely recognized as a critical determinant of disease progression, the precise spatiotemporal dynamics and mechanisms underlying vascular damage remain largely elusive, limiting the development of effective therapeutic interventions. Methods:An integrated imaging method that combines vascular labeling, tissue clearing, and light-sheet microscopy is proposed to realize 3D mapping and quantitative analysis of multi-organ vascular networks in a mouse model of SAP. Results:We found that SAP induced systemic microvascular disruption accompanied by organ-specific perfusion deficits across multiple vital organs. As the primary site of injury, the pancreas exhibited the earliest and most severe hypoperfusion, whereas remote organs showed a delayed decline in perfusion, with different trends among organs. Specifically, perfusion in the lung, heart, and kidney decreased gradually with disease progression, perfusion in the liver and spleen remained stable within the first 6 hours and then decreased rapidly, while the brain perfusion showed a sharp decrease at the end stage. These distinct perfusion trajectories are consistent with sequential compensatory and decompensatory responses. More importantly, the administration of heparin sodium within 12 hours after SAP induction significantly improved survival, reversed multi-organ hypoperfusion, and preserved microvascular integrity across affected organs. Conclusions:This study identifies systemic vascular injury as a key driver of multiple organ dysfunction in SAP and provides a convincing evidence that early intervention with heparin sodium can improve multi-organ hypoperfusion, preserve microvascular integrity, and enhance survival.
Background:Despite its promising therapeutic potential, inefficient site-specific targeting and complex fabrication processes hinder the clinical translation of oral melatonin therapy for ulcerative colitis (UC). Herein, we present Melatonin-Eudragit-Thioketal polymer-based Assembly (META), a dual pH/ROS-responsive oral microsphere for precise melatonin delivery to the inflamed colon and enhanced therapeutic outcomes. Methods:META was fabricated via a scalable emulsification process using Eudragit® FS 30 D and a thioketal polymer to enable dual pH- and ROS-responsive release. Its targeting delivery and therapeutic effects were then evaluated in simulated gastrointestinal (GI) fluids, 2D cell models, intestinal organoids, and a murine colitis model. Results:Using cell and animal models, we showed the important role of melatonin in GI health, supporting our strategy for UC treatment. META exhibited controlled melatonin release and better accumulation in the inflamed colon. The targeted delivery helped promote disease recovery by modulating macrophages and improving the intestinal barrier. Conclusion:META delivers melatonin precisely to the colon, which reprograms colonic macrophages and reinforces the epithelial barrier. This study offers a practical, clinically relevant approach for improving the use of endogenous therapeutic molecules in UC and other inflammatory GI diseases.
The prognosis of metastatic castration-resistant prostate cancer is dismal. [177Lu]Lutetium-prostate-specific membrane antigen (PSMA)-617 ([177Lu]Lu-PSMA-617) is the new standard of care for PSMA-positive mCRPC previously treated with ARPI and taxane-based chemotherapy. The efficacy and safety of [177Lu]Lu-PSMA-617 have been confirmed by phase 2 (TheraP [NCT03392428]) and phase 3 (VISION study [NCT02511664] and PSMAfore trial [NCT04689828]) clinical trials. A substantial proportion of patients with mCRPC are older adults; however, chronological age alone should not be considered a contraindication to [177Lu]Lu-PSMA-617 therapy. Instead, therapeutic decisions must be guided by a comprehensive evaluation of life expectancy, comorbidities, and frailty. Here we review the effectiveness and safety data for [177Lu]Lu-PSMA-617, present tools for the geriatric and frailty assessments in clinical practice and discuss its utility and therapeutic indications in patients with different types of frailty.
Rationale:[18F]meta-fluorobenzylguanidine ([18F]MFBG) is an human norepinephrine transporter (hNET)-targeted PET tracer that has shown superior lesion detection than [123I]MIBG SPECT in patients with relapsed or refractory neuroblastoma. Its value in newly diagnosed patients, however, has not been well established. We therefore performed this study to evaluate the additional staging information of [18F]MFBG beyond comprehensive anatomic imaging and assessed its agreement with [18F]FDG PET at baseline. Methods:We prospectively enrolled consecutive children with newly diagnosed neuroblastoma. All patients underwent baseline [18F]MFBG PET and comprehensive anatomic imaging, including contrast-enhanced CT and/ or MRI from the neck to the pelvis and whole-body ultrasonography. The imaging studies were interpreted independently, and lesion distribution and International Neuroblastoma Risk Classification (INRG) stage were recorded. A subgroup also underwent paired [18F]FDG PET for functional comparison. Paired detection rates were compared using the McNemar test. Intermodality concordance was evaluated using Cohen κ statistics, and Curie scores were compared using Wilcoxon signed-rank tests. Results:Forty patients (median age, 23.5 months) were analyzed. The primary tumors were mainly located in the abdomen or pelvis (36/40, 90%), with corresponding imaging findings including contrast-enhanced abdominopelvic CT and MRI, non-contrast cervicothoracic CT, and whole-body ultrasonography. [18F]MFBG PET identified bone marrow metastases more frequently than anatomic imaging (65% vs. 32.5%, P = 0.001; κ = 0.41), leading to stage reclassification in 20% of cases and treatment modification in 10% based on tumor-board review. Even within overlapping fields of view (neck to proximal femur), [18F]MFBG maintained superior bone marrow detection rates (55% vs. 32.5%, P = 0.004; κ = 0.56). Among the patients initially classified as L2, half were upstaged based on hNET-targeted PET findings. In the paired [18F]FDG PET subset (n = 26), the patient-level INRG stage was fully concordant between modalities. However, lesion-level analysis revealed a higher bone marrow disease burden on [18F]MFBG (191 vs. 140, P = 0.002), while FDG detected 13% additional nodal lesions in five patients. Conclusion:In newly diagnosed neuroblastoma, hNET-targeted [18F]MFBG PET detected clinically relevant bone marrow involvement not identified by comprehensive anatomic imaging, resulting in stage reclassification and treatment modification in a subset of patients. Paired [18F]FDG PET demonstrated patient-level stage concordance with lesion-level heterogeneity.
Surrogate materials enable pre-treatment prediction of intra-hepatic distribution of therapeutic radioactive microsphere, facilitating patient-specific dosimetry for transarterial radioembolization (TARE). Technetium-99m macroaggregated albumin (⁹⁹ᵐTc-MAA) has long constituted the clinical standard for SPECT-based surrogates, however, its fundamental physicochemical difference from therapeutic microspheres limits the reliability of MAA-based dosimetry and prediction for personalized treatment planning. Next-generation surrogate technology includes biodegradable polymeric microspheres engineered to closely replicate the therapeutic microsphere while affording post-procedural arterial recanalization through controlled biodegradation. Theranostic microspheres incorporating diverse radionuclides, positron-emitting PET compatible surrogates, and multimodal CT and MRI-visible surrogates have collectively expanded same-particle pre-treatment dosimetry capabilities. These material-level advances are synergistically coupled with computational dosimetry tools including artificial intelligence/deep learning algorithms for automated segmentation/dose-prediction, and Monte Carlo simulations that enhance voxel-based dosimetry precision. This review examines recent advances in surrogate materials design and integrated imaging/dosimetry methodologies that enhance TARE efficacy, safety, and personalized treatment outcomes.
Pathological cardiac hypertrophy is maladaptive cardiac remodeling induced by chronic adverse stimuli. In this study, the E3 ubiquitin ligase RNF128 was identified as a suppressor of pathological cardiac dysfunction with therapeutic value. Methods:The expression of Ring Finger protein 128 (RNF128) in pathological cardiac hypertrophy was characterized via public database analysis, scRNA-seq (single-cell RNA sequencing), and further validated in clinical myocardial samples and mouse disease models. The regulatory function of RNF128 in the progression of cardiac hypertrophy was verified in vivo by cardiomyocyte-specific RNF128 knockout mice and cTnT-AAV9-mediated RNF128 overexpression. Ang II (angiotensin II)-stimulated NMCMs (neonatal mouse cardiomyocytes) were used for in vitro validation. Moreover, the downstream target of RNF128 was identified through integrated analysis of scRNA-seq, interactome profiling and quantitative proteomics, followed by a panel of molecular assays. Results:Pathological cardiac hypertrophy reduced RNF128 expression in both human and murine samples. RNF128 deficiency aggravated cardiac dysfunction and pathological remodeling, while its overexpression protected cardiac function. Mechanistically, RNF128 directly interacted with SERCA2a, and catalyzed K63-linked polyubiquitination of SERCA2a at residue K476 (lysine 476), thereby impaired SERCA2a recognition by SQSTM1/p62. Consequently, RNF128 inhibited autophagy-lysosome-mediated degradation of SERCA2a. Conclusions:The findings of this study highlight RNF128 as a novel therapeutic target for heart failure, linking ubiquitination-dependent protein regulation to calcium handling in cardiomyocytes.
Rationale:Endometrial injury exhibits significant clinical heterogeneity in fibrotic outcomes. Recovery of microvascular perfusion is central to prognosis, while the depth of injury is an important factor influencing regenerative capacity. However, current clinical imaging techniques are limited by diffraction, resulting in insufficient resolution to reliably depict depth-dependent microvascular architecture of the endometrium, with consequent difficulty in evaluating injury depth or in making early prognostic distinctions. Methods:A depth-derived ultrasound localization microscopy (D-ULM) approach is introduced to capture depth-dependent microvascular features in the endometrium. First, we established a normal depth reference band from healthy rats to provide a physiological baseline for identifying pathological vascular changes. Next, in an endometrial injury rat model, we applied unsupervised clustering of early D-ULM features to identify microcirculation phenotypes. Subsequently, we evaluated the correlation between D-ULM features, D-ULM clusters, and late-stage fibrosis area as well as immunofluorescence markers of hypoxia and inflammation, with a view to tying early microvascular changes to eventual recovery or to fibrotic remodeling. Results:In healthy rats, depth-profile curves revealed a distinct transition zone between the deep and superficial layers, enabling the delineation of a quantifiable functional boundary of the endometrial microvasculature. Based on this depth-dependent layering, normal reference intervals were established for the seven D-ULM features. Among these, the reference interval for the vascular distribution center (com_depth_vessel) was 0.371-0.481. In injured cohort, unsupervised hierarchical clustering of day-3 D-ULM features identified three phenotypes: regenerative (Reg), inflammatory hyperperfusion (IH), and irreversible destruction (ID). The day-14 fibrosis area differed significantly among the phenotypes (P < 0.001), with the Reg phenotype showing the lowest fibrosis, while the IH and ID phenotypes exhibited markedly higher levels. Several D-ULM features were positively correlated with day-14 fibrosis, including com_depth_vessel (ρ = 0.707, q < 0.001), vessel_slope (ρ = 0.539, q = 0.017), vessel_auc (ρ = 0.811, q < 0.001), and mean_vessel_shallow (ρ = 0.846, q < 0.001); conversely, large_ratio_diff was inversely correlated (ρ = -0.568, q = 0.011). The IH phenotype exhibited elevated M1/M2 ratio, HIF-1α, and CD31 expression, whereas the ID phenotype showed increased M1/M2 ratio and HIF-1α but reduced CD31 expression by immunofluorescence. Conclusions:D-ULM enables depth-resolved imaging of endometrial microvascular functional gradients in a noninvasive manner. This approach provides an imaging framework for early stratification of injury phenotypes, and may guide personalized preventive strategies for patients at risk of postinjury fibrosis.
